Consequences of disrupting the gene that encodes α-glucosidase II in the N-linked oligosaccharide biosynthesis pathway ofDictyostelium discoideum

Freeze, Hudson H.; Lammertz, Marion; Iranfar, Negin; Fuller, Danny; Panneerselvam, K.; Loomis, William F.

doi:10.1002/(sici)1520-6408(1997)21:3<177::aid-dvg1>3.0.co;2-4

Cited by 10 publications

(1 citation statement)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…ModA is a glycosidase present in the ER which plays a key role in the maturation and transport of many lysosomal enzymes. In modA KO cells, the activity of many lysosomal enzymes is significantly reduced [10]. We generated modA KO cells as detailed in the Material Methods section and in Fig.…”

Section: Resultsmentioning

confidence: 99%

Antibacterial effectors inDictyostelium discoideum: specific activity against different bacterial species

Munoz-Ruiz,

Lamrabet,

Jauslin

et al. 2024

Preprint

View full text Add to dashboard Cite

Dictyostelium discoideumis a phagocytic amoeba continuously eating, killing and digesting bacteria. Previous studies have detected inD. discoideumcell extracts a bacteriolytic activity effective againstKlebsiella pneumoniaebacteria.In this study we characterized bacteriolytic activities found inD. discoideumcell extracts against five different bacteria (K. pneumoniae,Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus, andBacillus subtilis).We first analyzed the bacteriolytic activity against these five bacteria in parallel over a range of pH values. We then measured the remaining bacteriolytic activity inD. discoideum kil1andmodAKO mutants. We also performed partial fractionation ofD. discoideumextracts and assessed activity against different bacteria. Together our results indicate that optimal bacteriolytic activity against different bacteria results from the action of different effectors. Proteomic analysis allowed us to propose a list of potential bacteriolytic effectors.IMPORTANCEMany antibacterial effectors have been characterized over the last decades, and their biological importance, mode of action and specificity is often still under study. Here we characterizedin vitrobacteriolytic activity inD. discoideumextracts against five species of gram-negative and gram-positive bacteria. Our results reveal that optimal lysis of different bacteria mobilizes different effectors. Proteomic analysis generated a list of potential bacteriolytic effectors. This work opens the way for future analysis of the role of individual effectors in livingD. discoideumcells.

show abstract

Section: Resultsmentioning

confidence: 99%

Antibacterial effectors inDictyostelium discoideum: specific activity against different bacterial species

Munoz-Ruiz,

Lamrabet,

Jauslin

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

N‐glycomic profiling of a glucosidase II mutant of Dictyostelium discoideum by ‘‘off‐line’’ liquid chromatography and mass spectrometry

et al. 2014

View full text Add to dashboard Cite

In this study, we have performed the first mass spectrometric analysis of N-glycans of the M31 mutant strain of the cellular slime mould Dictyostelium discoideum, previously shown to have a defect in glucosidase II. Together with glucosidase I, this enzyme mediates part of the initial processing of N-glycans; defects in either glucosidase are associated with human diseases and result in an accumulation of incorrectly-processed oligosaccharides which are not, or only poor, substrates for a range of downstream enzymes. To examine the effect of the glucosidase II mutation in Dictyostelium, we employed off-line LC-MALDI-TOF-MS in combination with chemical and enzymatic treatments and MS/MS to analyse the neutral and anionic N-glycans of the mutant as compared to the wild-type. The major neutral species were, as expected, of the composition Hex 10-11 HexNAc 2-3 with one or two terminal glucose residues. Consistent with the block in processing of neutral N-glycans caused by the absence of glucosidase II, fucose was apparently absent from the N-glycans and bisecting N-acetylglucosamine was rare. The major anionic oligosaccharides were sulphated and/or methylphosphorylated forms of Hex 8-11 HexNAc 2-3 , many of which surprisingly lacked glucose residues entirely. As anionic Nglycans are considered to be mostly associated with lysosomal enzymes in Dictyostelium, we hypothesise that glycosidases present in the acidic compartments may act on the oligosaccharides attached to such slime mould proteins. Furthermore, our chosen analytical approach enabled us, via observation of diagnostic negative-mode MS/MS fragments, to determine the fine structure of the methylphosphorylated and sulphated N-glycans of the M31 glucosidase mutant in their native state.

show abstract

N-glycan trimming by glucosidase II is essential for Arabidopsis development

et al. 2008

View full text Add to dashboard Cite

Glucosidase II, one of the early N-glycan processing enzymes and a major player in the glycoprotein folding quality control, has been described as a soluble heterodimer composed of α and β subunits. Here we present the first characterization of a plant glucosidase II α subunit at the molecular level. Expression of the Arabidopsis α subunit restored N-glycan maturation capacity in Schizosaccharomyces pombe α-or αβ-deficient mutants, but with a lower efficiency in the last case. Inactivation of the α subunit in a temperature sensitive Arabidopsis mutant blocked N-glycan processing after a first trimming by glucosidase I and strongly affected seedling development.

show abstract

Consequences of disrupting the gene that encodes α-glucosidase II in the N-linked oligosaccharide biosynthesis pathway ofDictyostelium discoideum

Cited by 10 publications

References 53 publications

Antibacterial effectors inDictyostelium discoideum: specific activity against different bacterial species

Antibacterial effectors inDictyostelium discoideum: specific activity against different bacterial species

N‐glycomic profiling of a glucosidase II mutant of Dictyostelium discoideum by ‘‘off‐line’’ liquid chromatography and mass spectrometry

N-glycan trimming by glucosidase II is essential for Arabidopsis development

Contact Info

Product

Resources

About